Enhanced heterologous protein display on bacterial magnetic particles using a lon protease gene deletion mutant in Magnetospirillum magneticum AMB-1

Kanetsuki, Yuka; Tanaka, Tsuyoshi; Matsunaga, Tadashi; Yoshino, Tomoko

doi:10.1016/j.jbiosc.2013.01.016

Cited by 8 publications

(2 citation statements)

References 29 publications

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“…The subcellular localization analysis indicated that elimination of this putative transmembrane region resulted in the change in localization of the Mms6 protein. Alternatively, the Mms6 variants are thought to be digested by endogenous proteases prior to the localization of the protein to the proper position in the magnetosome membrane 48 because the His-tag fused proteins were also absent in the cytoplasm and cell membrane fractions. Moreover, elimination of the C-terminal region influenced the localization of proteins onto the surface of magnetite crystals.…”

Section: Discussionmentioning

confidence: 99%

Core Amino Acid Residues in the Morphology-Regulating Protein, Mms6, for Intracellular Magnetite Biomineralization

Yamagishi

Narumiya

Tanaka

et al. 2016

Sci Rep

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Living organisms produce finely tuned biomineral architectures with the aid of biomineral-associated proteins. The functional amino acid residues in these proteins have been previously identified using in vitro and in silico experimentation in different biomineralization systems. However, the investigation in living organisms is limited owing to the difficulty in establishing appropriate genetic techniques. Mms6 protein, isolated from the surface of magnetite crystals synthesized in magnetotactic bacteria, was shown to play a key role in the regulation of crystal morphology. In this study, we have demonstrated a defect in the specific region or substituted acidic amino acid residues in the Mms6 protein for observing their effect on magnetite biomineralization in vivo. Analysis of the gene deletion mutants and transformants of Magnetospirillum magneticum AMB-1 expressing partially truncated Mms6 protein revealed that deletions in the N-terminal or C-terminal regions disrupted proper protein localization to the magnetite surface, resulting in a change in the crystal morphology. Moreover, single amino acid substitutions at Asp123, Glu124, or Glu125 in the C-terminal region of Mms6 clearly indicated that these amino acid residues had a direct impact on magnetite crystal morphology. Thus, these consecutive acidic amino acid residues were found to be core residues regulating magnetite crystal morphology.

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Section: Discussionmentioning

confidence: 99%

Core Amino Acid Residues in the Morphology-Regulating Protein, Mms6, for Intracellular Magnetite Biomineralization

Yamagishi

Narumiya

Tanaka

et al. 2016

Sci Rep

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“…Therefore, the lower binding activity may also be explained by either lack of glycosylation or the resulting partially incorrect folding. Strategies to increase the expression level will include optimization of induction conditions [13], selection of an appropriate promoter [27] and/or genetic modification of the host strain [28,29]. In addition, it may be helpful to test the effect on ligand binding activity of lower growth temperatures, co-expression of molecular chaperones [30] or the formation of disulfide bonds [31].…”

Section: Discussionmentioning

confidence: 99%

Functional Expression of Thyroid-Stimulating Hormone Receptor on Nano-Sized Bacterial Magnetic Particles in Magnetospirillum magneticum AMB-1

Sugamata

Uchiyama

Honda

et al. 2013

IJMS

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The measurement of autoantibodies to thyroid-stimulating hormone receptor (TSHR) is important for the diagnosis of autoimmune thyroid disease such as Graves’ disease (GD). Although TSHR from porcine thyroid membrane is commonly used for the measurement of TSHR autoantibodies (TRAb), recombinant human TSHR (hTSHR) remains ideal in terms of stable supply and species identity. Here we set out to express recombinant hTSHR on the lipid-bilayer surface of magnetic nanoparticles from a magnetotactic bacterium, Magnetospirillum magneticum AMB-1. Using a tetracycline-inducible expression system, we successfully overexpressed functional hTSHR on bacterial magnetic particles (BacMPs) in AMB-1 via an anchor protein specific for BacMPs. The overexpressed hTSHR was membrane integrated and possessed both ligand and autoantibody binding activity. Our data suggest that hTSHR-displayed BacMPs have potential as novel tools for ligand-receptor interaction analysis or for TRAb immunoassay in GD patients.

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Magnetotactic Bacteria, Magnetosomes, and Nanotechnology

2014

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Enhanced heterologous protein display on bacterial magnetic particles using a lon protease gene deletion mutant in Magnetospirillum magneticum AMB-1

Cited by 8 publications

References 29 publications

Core Amino Acid Residues in the Morphology-Regulating Protein, Mms6, for Intracellular Magnetite Biomineralization

Core Amino Acid Residues in the Morphology-Regulating Protein, Mms6, for Intracellular Magnetite Biomineralization

Functional Expression of Thyroid-Stimulating Hormone Receptor on Nano-Sized Bacterial Magnetic Particles in Magnetospirillum magneticum AMB-1

Magnetotactic Bacteria, Magnetosomes, and Nanotechnology

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